WO2017199755A1 - Drug for preventing and/or treating dementia - Google Patents

Abstract

Provided is a drug that has excellent effect of preventing and/or treating dementia. The drug comprises a carbostyril derivative represented by formula (1) [wherein: R represents a cycloalkyl group; A represents a lower alkyl group; and the bond between the 3- and 4-positions of the carbostyril nucleus is either a single bond or a double bond] and dihydroquercetin.

Description

Drugs for prevention and / or treatment of dementia

The present invention relates to a drug for preventing and / or treating dementia.

Dementia refers to a state in which memory function and other cognitive functions have deteriorated to such an extent that problems in daily life occur due to changes in the brain. The main causative diseases of dementia include Alzheimer's dementia and vascular dementia. In dementia, there are memory disorders, disorientation, executive dysfunction, handwriting and calculation disorders, which are called core symptoms, and disorders such as anxiety, sleeplessness, increased aggression, depression, depression, and delirium, which are called peripheral symptoms. There is.

Memory impairment and disorientation in human dementia patients correspond to spatial memory impairment in dementia model animals. Spatial memory means the ability to grasp and memorize the state and relationship that an object occupies in a three-dimensional space, such as the position, orientation, posture, size, shape, and spacing of an object. It means such a disorder of spatial memory. Further, the executive function failure is also referred to as working memory failure. Working memory means the ability to keep information in mind and process it simultaneously in a short time, and working memory failure means such working memory failure.

“Prevention and treatment of dementia is important not only in improving the patient's QOL, but also in reducing the burden on the family due to care and reducing medical costs. The main causative diseases of dementia in Japan include Alzheimer's dementia with cerebral amyloid plaques and vascular dementia caused by cerebrovascular disease. In particular, the former accounts for nearly half of all dementia patients. Has been attracting attention.

However, in recent years, it has been suggested that vascular pathology is also deeply involved in the pathogenesis of Alzheimer's disease, and in the treatment of Alzheimer's disease, an approach from the cardiovascular side is being sought.

An example of this is cilostazol. Cilostazol is an antiplatelet agent that is used as a preventive agent for recurrence after cerebral infarction such as lacunar infarction. That is, by taking cilostazol, accumulation of (i) amyloid beta (hereinafter sometimes abbreviated as Aβ) in the brain is suppressed (Non-patent Documents 1 and 2), and (ii) cognitive function decrease is suppressed. (Non-patent Documents 1 and 2), and (iii) the effect of increasing cerebral blood flow in the vertebral artery, internal carotid artery, cerebral cortex, hypothalamus, etc. (cilostazol package insert) has been reported. Regarding (i), it is said that the activation of the perivascular drainage pathway (brain interstitial flow), which is a waste discharge system in the brain, is involved.

Peri-cerebrovascular drainage pathway is likely to be a universal removal route for not only Aβ but also neurotoxic substances such as tau protein and α-synuclein protein, and oral cilostazol causes various cognitive impairments caused by accumulation of many neurotoxic proteins. It is expected to be effective against (eg, Alzheimer's disease, frontotemporal lobar degeneration, dementia with Lewy bodies, etc.). Although cerebral blood flow reduction / cerebrovascular disorder is a factor that promotes the onset and progression of dementia, oral cilostazol also has a clear effect in both cerebral blood flow improvement and cerebral infarction prevention. Yes.

In Non-Patent Document 3, as a result of administration of cilostazol-containing food to APPSwDI genetically modified mice, APPSwDI genetically modified mice administered with cilostazol had a Y-type maze compared to APPSwDI genetically modified mice administered with control food. It has been described that the alternation response in the trial has increased, suggesting that cilostazol is beneficial in improving working memory impairment in dementia.

The present invention aims to further improve the effect of a drug for the prevention and / or treatment of dementia including cilostazol. Specifically, the present invention aims to provide an effective drug that is effective not only in the core symptoms of dementia but also in peripheral symptoms, and even in a state in which the symptoms have progressed.

The agent for preventing and / or treating dementia according to the present invention is a carbostyril derivative having the following formula (1) (wherein R is a cycloalkyl group, A is a lower alkyl group, carbostyril). Between the 3rd and 4th positions of the nucleus is a single bond or a double bond) or a salt thereof,

 

And dihydroquercetin or a salt thereof.

The agent for prevention and / or treatment of dementia according to the present invention is a carbostyril derivative comprising the following formula (1) as an active ingredient (where R is a cycloalkyl group and A is a lower alkyl group). And between the 3rd and 4th positions of the carbostyryl nucleus is a single bond or a double bond) or a salt thereof,

 

An active ingredient dihydroquercetin or a salt thereof;
Is administered in combination.

According to the present invention, further prevention and / or treatment effects of dementia can be obtained as compared with the case of cilostazol alone.

It is a figure which shows the amount of A (beta) deposition to the cerebral blood-vessel wall in the mouse | mouth of each group. In mice in each group is a diagram showing an average value of 5% CO ₂ relative increase rate of cerebral blood flow after air. It is a figure which shows the external appearance of the circular pool used by a water maze test, Among them, (A) is a cross-sectional view, (B) is a top view. It is a figure which shows the result of a water maze test, (A) is a figure which shows the average value of the total swimming distance of the mouse | mouth of each group, (B) is a figure which shows the average value of the swimming time of the mouse | mouth of each group. (C) is a figure which shows the average value of the residence time of Zone-1 of the mouse | mouth of each group in a probe test. It is a figure which shows the cumulative survival rate of the mouse | mouth of each group. It is a figure which shows the average value of the hair loss score of the mouse | mouth of each group. It is a figure which shows the result of a Y maze test.

Hereinafter, embodiments of the present invention will be specifically described with reference to the accompanying drawings. However, the embodiments are for facilitating understanding of the principle of the present invention, and the scope of the present invention is as follows. The present invention is not limited to the embodiments, and other embodiments in which those skilled in the art appropriately replace the configurations of the following embodiments are also included in the scope of the present invention.

The agent for preventing and / or treating dementia according to the present embodiment is a carbostyril derivative having the following formula (1) (wherein R is a cycloalkyl group, A is a lower alkyl group, Between the 3- and 4-positions of the styryl nucleus is a single bond or a double bond) or a salt thereof,

 

Dihydroquercetin or a salt thereof.

The agent for preventing and / or treating dementia according to the present embodiment is a carbostyril derivative comprising the following formula (1) as an active ingredient (where R is a cycloalkyl group, and A is a lower alkyl group). And between the 3rd and 4th positions of the carbostyryl nucleus is a single bond or a double bond) or a salt thereof,

 

An active ingredient dihydroquercetin or a salt thereof;
In combination.

In the above formula (1), the cycloalkyl group includes, for example, C ₃ -C ₈ cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. A preferred cycloalkyl group is cyclohexyl.

In the above formula (1), the lower alkyl group includes, for example, C ₁ -C ₆ alkylene groups such as methylene, ethylene, propylene, tetramethylene, butylene and pentylene, and tetramethylene is preferred.

The drug according to this embodiment is for the prevention and / or treatment of dementia. In this specification, “prevention” includes suppressing and delaying the onset of the disease, resulting in the disease. This includes not only pre-prevention but also prevention of disease recurrence after treatment. On the other hand, “treatment” includes curing a symptom, improving the symptom, and suppressing the progression of the symptom. Dementia is not particularly limited, for example, Alzheimer's dementia, Lewy body dementia, frontotemporal dementia, cerebrovascular dementia, Parkinson's disease, Down's syndrome, or Huntington's disease Is included. The drug according to this embodiment is for the prevention and / or treatment of dementia, but also enables the prevention and / or treatment of not only the core symptoms of dementia but also the peripheral symptoms that accompany the core symptoms. . A peripheral symptom is a symptom that appears frequently as dementia progresses from moderate to severe. Peripheral symptoms include aggressive behavior, delusions, sleep disturbances, jealousy, resistance to care, falls due to hyperactivity, suffocation due to impulsive stealing, and the like.

The drug according to the present embodiment has a carbostyril derivative or a salt thereof and dihydroquercetin or a salt thereof, but includes a form in which these combinations are administered simultaneously or separately or sequentially. For example, a tablet or fine granule containing a carbostyril derivative as an active ingredient and a tablet or fine granule containing dihydroquercetin as an active ingredient are combined, and these are administered sequentially. Are included in such drugs.
In other words, “the drug having carbostyril derivative or a salt thereof and dihydroquercetin or a salt thereof” corresponds to, for example, the following forms.
A form in which the active ingredient carbostyril derivative or a salt thereof and the active ingredient dihydroquercetin or a salt thereof are administered simultaneously or separately or sequentially.
A form containing an active ingredient carbostyril derivative or a salt thereof and an active ingredient dihydroquercetin or a salt thereof.

A preferred carbostyril derivative is 6- [4- (1-cyclohexyl-1H-tetrazol-5-yl) butoxy] -3,4-dihydrocarbostyril represented by the following formula (2), and cilostazol as an antiplatelet agent It is on the market with the product name.

 

The average particle size of cilostazol is not particularly limited, but is preferably 10 μm to 2000 μm, for example. This is because if the average particle size is larger than 2000 μm, an expensive apparatus is required for preparing the resin particles, and if the average particle size is smaller than 10 μm, the absorption in the lower digestive tract may be deteriorated. Because there is.

The carbostyril derivative can easily form a salt by acting a pharmaceutically acceptable acid. Examples of the acid include inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, and hydrobromic acid, and organic acids such as oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, and benzoic acid. .

A carbostyril derivative and a salt thereof, and a production method thereof are disclosed in JP-A-55-35019 (corresponding US Pat. No. 4,277,479).

Dihydroquercetin is a compound represented by the following formula (3).

 

Dihydroquercetin has four types of stereoisomers depending on the configuration at the 2nd and 3rd positions. One of the compounds represented by the following formula (4) is called taxifolin.

 

Dihydroquercetin includes dihydroquercetin derivatives. Examples of the dihydroquercetin derivative include glycosides in which a sugar is bonded to at least one hydroxyl group at the 3, 3 ', 4', 5 or 7 position of dihydroquercetin.

As a salt of dihydroquercetin, a salt can be easily formed by acting a pharmaceutically acceptable acid. Examples of such salts include alkali metal salts such as sodium salts and potassium salts.

Dihydroquercetin or a salt thereof can be produced, for example, by chemical synthesis or extraction from Siberian larch.

The blending ratio of the carbostyril derivative and dihydroquercetin is not particularly limited, but may be, for example, 1: 5 to 1:20 by weight.

The dose of the active ingredient in the drug according to the present embodiment can be appropriately set depending on the age, sex, weight, symptoms, etc. of the patient. For example, in adults (weight 50 kg), 35 per day per carbostyril derivative. It is possible to administer ~ 400 mg, preferably 100 to 200 mg, and 150 to 2000 mg, preferably 500 to 1000 mg per day per dihydroquercetin, once or in two or several divided doses.

The administration method of the drug according to the present embodiment is not particularly limited. For example, the combination of carbostyril derivative and dihydroquercetin is administered simultaneously or separately, or sequentially with a time difference of several hours to several days. It is possible to administer the administration method. When administered sequentially, either component may be administered first.

The drug according to this embodiment is, for example, a preparation for oral administration such as tablets, granules, fine granules, capsules, various liquid preparations suitable for oral administration, or non-injections such as injections and suppositories. It is possible to prepare a preparation for oral administration.

In the case of a preparation for oral administration, it is obtained by formulating the fine powder of the drug according to the present embodiment and a dispersant and / or a dissolution improving agent in the form of tablets, granules, fine granules, capsules and the like together with a preparation carrier. By adding a dispersant and / or a dissolution improver, the dispersibility and / or dissolution absorbability of the fine powder of carbostyril derivative can be enhanced.

As the formulation carrier, excipients, binders, disintegrants, lubricants, plasticizers, and the like can be used. As the excipient, for example, sucrose, sodium chloride, mannitol, lactose, glucose, starch, calcium carbonate, face forest, crystalline cellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose acetate succinate, silicate, etc. are used. it can. As the binder, for example, water, ethanol, propanol, glucose solution, starch solution, gelatin solution, sodium carboxymethylcellulose, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone and the like can be used. As the disintegrant, for example, carboxymethylcellulose calcium, dry starch, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, stearic acid monoglyceride, starch, sodium carboxymethyl starch, croscarmellose sodium and the like can be used. As the lubricant, for example, purified talc, stearate, boric acid powder, polyethylene glycol, colloidal silicic acid, hydrogenated oil and the like can be used. As the plasticizer, for example, glycerin fatty acid ester, dioctyl phthalate, dibutyl phthalate, triacetin, triethyl citrate, castor oil and the like can be used.

As a dispersing agent and / or a dissolution improving agent, a water-soluble polymer and a surfactant can be used. As the water-soluble polymer, for example, hydroxypropylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, hydroxypropylmethylcellulose, carboxymethylcellulose, polyacrylic acid and the like can be used. Examples of the surfactant include alkyl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; polyglycerin fatty acid esters such as decaglyceryl monolaurate and decaglyceryl monomyristate; polyoxyethylene such as polyoxyethylene sorbitan monooleate Sorbitan fatty acid ester; Polyethylene glycol fatty acid ester such as polyoxyethylene monostearate; Polyoxyethylene alkyl ether such as polyoxyethylene lauryl ether; Polyoxyethylene castor oil such as polyoxyethylene hydrogenated castor oil and hydrogenated castor oil; Sucrose Sucrose fatty acid esters such as stearic acid ester and sucrose barmitic acid ester can be used.

It is preferable to add 0.001 to 100 parts by weight, preferably 0.01 to 10 parts by weight of the dispersant and / or dissolution improver to 1 part by weight of the fine powder of the drug according to this embodiment. Absorption decreases when the amount of the dispersant and / or dissolution improver is less than 0.001 part by weight, whereas when the amount is greater than 100 parts by weight, it depends on toxicity such as mucosal disorder or the Pharmaceutical Affairs Law. This is because there is a possibility of being restricted.

To prepare a tablet, the drug according to this embodiment is made into a tablet by a conventional method using the above-mentioned preparation carrier. Granules or fine granules are prepared by adding the above-mentioned preparation carrier to the fine powder of the drug according to this embodiment, fluidized bed granulation, high speed stirring granulation, stirring fluidized bed granulation, centrifugal fluidized granulation, extrusion granulation, etc. It can be prepared by granulating. Capsules are prepared by mixing with an inert pharmaceutical filler or diluent and packed into hard gelatin capsules or soft capsules.

The adjustment of the average particle diameter of the drug according to the present embodiment can be formed using, for example, a hammer mill, a jet mill, a rotating ball mill, a vibration ball mill, a shaker mill, a rod mill, a tube mill, or the like.

The drug according to the present embodiment can be coated with a sustained-release coating base on tablets, granules, and fine granules. As the sustained-release coating base, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid copolymer, ethylcellulose and the like can be used. Thereby, for example, it is possible to provide a drug elution ability in the lower part of the digestive tract.

The oral liquid preparation is prepared by mixing the drug according to the present embodiment with a sweetener (for example, sucrose), a preservative (for example, methylparaben, propylparaben), a coloring agent, a fragrance, and the like.

Among the preparations for parenteral administration, preparations for injection are prepared, for example, in the form of solutions, emulsions or suspensions, and are made isotonic with blood. Formulations in the form of liquids, emulsions or suspensions are prepared using, for example, an aqueous medium, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid ester . Examples of the aqueous medium include water or a medium containing water. As water, sterilized water is used. Examples of the medium containing water include physiological saline, PBS (phosphate buffered physiological saline), lactic acid-containing Ringer's solution, and the like.

In the preparation for injection, additives usually used in the art can be appropriately used. Examples of the additive include isotonic agents, stabilizers, buffers, preservatives, chelating agents, antioxidants, and solubilizing agents. Examples of the isotonic agent include sugars such as glucose, sorbitol and mannitol, sodium chloride, glycerin, propylene glycol, polyethylene glycol and the like. Examples of the stabilizer include sodium sulfite. Examples of the buffer include borate buffer, phosphate buffer, citrate buffer, tartaric acid buffer, and acetate buffer. Examples of the preservative include paraoxybenzoic acid ester, benzyl alcohol, chlorocresol, phenethyl alcohol, benzethonium chloride and the like. Examples of chelating agents include sodium edetate and sodium citrate. Examples of the antioxidant include sodium sulfite, sodium hydrogen sulfite, sodium ascorbate, sodium thiosulfate and the like. Examples of the solubilizer include dextran, polyvinylpyrrolidone, sodium benzoate, ethylenediamine, salicylic acid amide, nicotinic acid amide, polyoxyethylene hydrogenated castor oil derivative, and the like.

The pH preparation may be contained in the injectable preparation. The pH adjuster may be an acid or a base. Specifically, examples of the acids include ascorbic acid, hydrochloric acid, gluconic acid, acetic acid, lactic acid, boric acid, phosphoric acid, sulfuric acid, tartaric acid, and citric acid. Examples of the base include potassium hydroxide, calcium hydroxide, sodium hydroxide, magnesium hydroxide, monoethanolamine, diethanolamine, triethanolamine and the like.

In addition to the human, the drug according to this embodiment can be applied to mammals other than humans such as monkeys, cows, horses, pigs, sheep, dogs, cats, rats, and mice.

The drug according to this embodiment has a remarkable effect due to the combination of a carbostyril derivative and dihydroquercetin. That is, as shown in the examples described later, cilostazol is effective for maintaining working memory, but may not be effective for maintaining spatial memory, but it is also effective to maintain spatial memory by using taxifolin together. . On the other hand, excision (fighting, etc.) may occur when taxifolin is administered alone, but excitement is sedated by the combined use of cilostazol. That is, when taxifolin alone is administered, the onset of peripheral symptoms that are difficult to avoid can be effectively prevented, or the suppressive action or improvement action on the peripheral symptoms after the onset can be expected. For patients with dementia, antipsychotics, mood stabilizers, anti-anxiety drugs, hypnotics, etc. may be used to suppress peripheral symptoms, but according to the drug according to this embodiment, dementia Therefore, it can be expected to reduce the types and amounts of medicines taken by patients.

As described above, the drug according to the present embodiment is effective not only for the core symptoms of dementia typified by memory impairment, but also for peripheral symptoms such as increased aggression and anxiety, and is extremely useful for treating dementia. It is valid.

1. Immunohistochemistry 8-month-old normal diet-administered APPSwDI gene-modified mice (n = 5), cilostazol-containing diet-administered APPSwDI gene-modified mice (n = 5), taxifolin-containing diet-administered APPSwDI gene-modified mice (n = 6), cilostazol and Analysis was performed on a total of 22 APPSwDI gene-modified mice (n = 6) administered with taxifolin-containing food (drug-containing food according to this example). Cilostazol-containing food, taxifolin-containing food, and cilostazol and taxifolin-containing food were administered from 4 weeks of age to 8 months of age. The cilostazol concentration in the cilostazol-containing diet was 0.3 wt%, and the taxifolin concentration in the taxifolin-containing diet was 3 wt%. Cilostazol and taxifolin concentrations in the cilostazol and taxifolin-containing diets were 0.3 wt% and 3 wt%, respectively. All APPSwDI gene-modified mice were homozygous male mice. It should be noted that the 4-week-old APPSwDI gene-modified mouse is considered to be an early stage where Aβ accumulates in the cerebral blood vessels, that is, an early stage of cerebral amyloid angiopathy, and necrosis of neurons in the brain is considered to be a stage where it has not progressed so much. .

The brain of 8-month-old APPSwDI genetically modified mice was fixed by perfusion using 4% paraformaldehyde, and the removed brain was dehydrated over 1 day. Then, a paraffin block of the fixed brain tissue was prepared, and the paraffin block was prepared. The preparation was prepared by slicing the sample with a microtome at 6 microns, and Aβ deposited on the blood vessel wall was observed with a microscope by immunohistochemistry for Aβ. The hippocampal region was traced with a tissue section 1 mm outside from the median part, and the ratio of the Aβ accumulation area in the region of interest was measured using image analysis software ImageJ (National Institutes of Health. USA).

FIG. 1 shows the average value of the amount of Aβ deposited in each group. Error bars represent standard deviation.

Cilostazol-containing diet-administered APPSwDI gene-modified mice, taxifolin-containing diet-administered APPSwDI gene-modified mice, and cilostazol- and taxifolin-containing diet-administered APPSwDI gene-modified mice significantly increased brain Aβ accumulation compared with normal diet-administered APPSwDI gene-modified mice The amount was decreasing.
2. Carbon dioxide inhalation test 12-month-old normal diet-administered C57BL / 6J mice (n = 4), normal diet-administered APPSwDI gene-modified mice (n = 10), cilostazol-containing diet-administered APPSwDI gene-modified mice (n = 4), taxifolin Analysis was performed on a total of 30 diet-administered APPSwDI gene-modified mice (n = 5) and cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice (n = 7). Cilostazol, taxifolin, or a diet containing cilostazol and taxifolin was administered from 4 weeks to 12 months of age. The cilostazol concentration in the cilostazol-containing diet was 0.3 wt%, and the taxifolin concentration in the taxifolin-containing diet was 3 wt%. All APPSwDI gene-modified mice were homozygous male mice.

Mice were fixed by intraperitoneal injection of α-chloralose and urethane. Endotracheal intubation was performed and baseline cerebral blood flow was measured. Thereafter, 5% CO ₂ was supplied, and cerebral blood flow was measured over time for 5 minutes. The relative rate of increase was measured from the baseline and changes in cerebral blood flow after 5% CO ₂ insufflation. Cerebral blood flow was measured with a laser speckle blood flow meter (OZ-2, Omega Wave Inc.).

FIG. 2 is a graph showing the average value of the relative increase rate of cerebral blood flow after 5% CO ₂ insufflation in each group of mice. The horizontal axis represents the time (seconds) after the start of 5% CO ₂ insufflation. White circle group is C57BL / 6J mice with normal diet, square group is APPSwDI gene-modified mice with normal diet, triangle group is cilostazol-containing diet-administered APPSwDI gene-modified mice, diamond group is taxiphorin-containing diet-administered APPSwDI gene-modified mice, black circle group Shows cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice. Error bars represent standard error.

Compared with C57BL / 6J mice, normal diet-administered APPSwDI gene-modified mice had a decreased rate of increase in cerebral blood flow after 5% CO ₂ insufflation. In the cilostazol-containing diet-administered APPSwDI gene-modified mice and the taxifolin-containing diet-administered APPSwDI gene-modified mice, the rate of increase in cerebral blood flow was higher than that in the normal diet-administered APPSwDI gene-modified mice. In taxifolin containing feed administration APPSwDI genetically modified mice compared to rapidly increase cerebral blood flow increase from 5% CO ₂ After air, 5% CO ₂ After air in the cilostazol-containing feed administration APPSwDI genetically modified mice, The rate of increase in cerebral blood flow increased after a certain period of time. These results indicate that taxifolin and cilostazol improved abnormalities of vascular reactivity in APPSwDI transgenic mice by different mechanisms. However, in repeated measures analysis of variance, there was no significant difference between the normal diet-administered APPSwDI gene-modified mice group and the taxifolin-administered group and cilostazol-administered group, and the normal diet-administered APPSwDI gene-modified mice group and cilostazol and taxifolin-containing diet-administered APPSwDI gene modification Significant differences were only found between the mouse groups (p <0.001). It was suggested that the combined use of drugs with different mechanisms of action of pharmacological effects increases the therapeutic effect of dementia due to the expression of the action of each drug.
3. Water maze test 8 months old C57BL / 6J mice (n = 15), normal diet-administered APPSwDI gene-modified mice (n = 17), cilostazol-containing diet-administered APPSwDI gene-modified mice (n = 10), taxifolin-containing diet-administered APPSwDI gene A water maze test was performed on 61 mice (n = 10) and APPSwDI gene-modified mice (n = 9) administered with cilostazol and taxifolin-containing diet. Cilostazol-containing food, taxifolin-containing food, and cilostazol and taxifolin-containing food were administered from 4 weeks to 8 months of age. The cilostazol concentration in the cilostazol-containing diet was 0.3 wt%, and the taxifolin concentration in the taxifolin-containing diet was 3 wt%. Cilostazol and taxifolin concentrations in the cilostazol and taxifolin-containing diets were 0.3 wt% and 3 wt%, respectively. All APPSwDI gene-modified mice were homozygous male mice.

The water maze test is a test that evaluates the visual space cognitive ability of a mouse using the habit of avoiding water and trying to escape. The water maze used in this experiment was a Morris type water maze manufactured by Brain Science Idea, Inc., and a circular pool with an inner diameter of 120 cm and a wall height of 30 cm painted in black was used. A circular circular escape platform made of clear acrylic with a diameter of 10 cm and a height of 10 cm was set at 30 cm from the center of the pool and 30 cm from the periphery. In order for the mouse to memorize the various spatial arrangements around it, posters, photographs and other clues were placed on the wall, and the location of these clues was always constant during the experiment. Water was applied to the pool to a depth of 11 cm, and a platform was installed approximately 1 cm below the water surface as shown in FIG.

In all tests, Zone-2 in FIG. 3B was set as the start position. Zone-1 to Zone-4 indicate the four divided areas of the circular pool. The mouse was gently placed on the surface with the mouse facing the pool wall so that the platform position was not visible.

The test was conducted 4 times a day from the 1st day to the 4th day. At each implementation, the total swimming distance and the time to reach the platform (swimming time) were measured. If the platform was not reached on the platform within 60 seconds, the experimenter finished by placing the mouse on the platform for 15 seconds, and 60 seconds was the swimming time of the mouse.

On the fifth day, mice were allowed to swim for 60 seconds with the platform removed from the pool (probe test). At this time, the stay time of Zone-1 where the goal was set from the first day to the fourth day was measured.

FIG. 4 (A) shows the average value of the total swimming distance of the mice in each group from the first day to the fourth day, and FIG. 4 (B) shows the respective values from the first day to the fourth day. The average value of the time (swimming time) to reach the platform of the mice of the group is shown. In either case, the horizontal axis is the number of trials. White circle group is C57BL / 6J mice with normal diet, square group APPSwDI gene-modified mice with normal diet, triangular group is cilostazol-containing diet-administered APPSwDI gene-modified mice, diamond group is taxiphorin-containing diet-administered APPSwDI gene-modified mice, black circle group Shows cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice. Error bars represent standard error.

∙ There is no significant difference in total swimming distance among the 5 groups of mice, indicating that there is no significant difference in motor function of the mice. On the other hand, in the swimming time to reach the goal, C57BL / 6J mice shorten the time to reach the platform each time it is performed, but normal diet-administered APPSwDI genetically modified mice do not shorten, and visuospatial memory impairment It was suggested. This abnormality was similar in cilostazol-containing diet-administered APPSwDI gene-modified mice, and it was thought that cilostazol had a limited effect on the improvement of visuospatial memory impairment in APPSwDI gene-modified mice. On the other hand, the taxifolin-containing diet-administered APPSwDI gene-modified mice and cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice showed the same results as C57BL / 6J mice. Thereby, it was shown that the medicine concerning a present example improves visuospatial memory disorder of an APPSwDI gene modification mouse. Taxifolin was also shown to dramatically improve visuospatial memory impairment in APPSwDI genetically modified mice.

FIG. 4 (C) shows the average value (in seconds) of the time spent staying in Zone-1 of each group of mice in the probe test in which the goal that had been installed was removed. Error bars represent standard error.

Compared to C57BL / 6J mice, normal-administered APPSwDI gene-modified mice had a shorter residence time of Zone-1, which originally had the goal, suggesting impaired visual spatial memory. This abnormality was similar in cilostazol-containing diet-administered APPSwDI gene-modified mice, and it was thought that cilostazol had a limited effect on the improvement of visuospatial memory impairment in APPSwDI gene-modified mice. On the other hand, the taxifolin-containing diet-administered APPSwDI gene-modified mice and cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice showed the same results as C57BL / 6J mice. Thereby, it was shown that the medicine concerning a present example improves visuospatial memory disorder of an APPSwDI gene modification mouse. Taxifolin was also shown to dramatically improve visuospatial memory impairment in APPSwDI genetically modified mice.
4). Survival curve Normal-feeding APPSwDI genetically modified mice (n = 40) born from April 1, 2014 to November 30, 2015, cilostazol-containing diet-administered APPSwDI genetically-modified mice (n = 65), taxifolin-containing diet-administered APPSwDI Survival curves of mice were prepared and analyzed for genetically modified mice (n = 33) and cilostazol and taxifolin-containing diet-administered APPSwDI genetically modified mice (n = 33). Mice deaths do not include sacrifices associated with conducting animal experiments.

FIG. 5 shows the cumulative survival rate of mice in each group on the vertical axis and the number of days after birth on the horizontal axis.
A shows a group of normal diet-administered APPSwDI genetically modified mice, B shows a cilostazol-containing diet-administered APPSwDI gene-modified mouse group, C shows a taxifolin-containing diet-administered APPSwDI gene-modified mouse group, and D shows a dietary administration APPSwDI gene-modified mouse group containing cilostazol and taxifolin ing.

Normally, the life span of a mouse is 2 years or longer, and it is relatively rare for a mouse to die within 500 days of observation. However, the taxifolin-containing diet-administered APPSwDI gene-modified mice clearly had more deaths than other mice. On the other hand, surprisingly, no death of mice was observed in APPSwDI gene-modified mice administered with diet containing cilostazol and taxifolin. Thereby, it was shown that the medicine concerning this execution is effective also in peripheral symptoms, such as an aggressiveness increase and anxiety.
5). Hair loss score When mice fight, they experience hair loss. Therefore, the degree of hair loss of each mouse was described in four stages from Grade 0 to 3, and used as a hair loss score. The almost normal state was Grade 0, and the state where hair loss was observed throughout the whole body was Grade 3. It was classified into Grade: 1 and Grade: 2 in less than 50% and more than 50% of the whole table.

13-month-old C57BL / 6J mice (n = 5), normal diet-administered APPSwDI gene-modified mice (n = 14), taxifolin-containing diet-administered APPSwDI gene-modified mice (n = 4), cilostazol and taxifolin-containing diet-administered APPSwDI gene modification For analysis with a total of 35 mice (n = 12), with the mouse information concealed, two evaluators independently evaluated the hair loss score, and the average value of each evaluation was used as the hair loss score for the mouse. did.

FIG. 6 shows the average value of the hair loss score of each group of mice. Error bars represent standard error.

Taxiphorin-containing diet-administered APPSwDI gene-modified mice clearly had severe hair loss compared to normal diet-administered APPSwDI gene-modified mice. However, such a tendency was not observed in APPSwDI gene-modified mice fed with cilostazol and taxifolin, suggesting a decrease in fighting. Thereby, it was shown that the medicine concerning this execution is effective also in peripheral symptoms, such as an aggressiveness increase and anxiety.
6). Y maze test 13-month-old C57BL / 6J mice (n = 4), normal diet-administered APPSwDI gene-modified mice (n = 14), taxifolin-containing diet-administered APPSwDI gene-modified mice (n = 7), cilostazol and taxifolin-containing diet A total of 39 APPSwDI gene-modified mice (n = 14) were subjected to the Y maze test. Cilostazol-containing food, taxifolin-containing food, and cilostazol and taxifolin-containing food were administered from 4 weeks to 13 months of age. The cilostazol concentration in the cilostazol-containing diet was 0.3 wt%, and the taxifolin concentration in the taxifolin-containing diet was 3 wt%. Cilostazol and taxifolin concentrations in the cilostazol and taxifolin-containing diets were 0.3 wt% and 3 wt%, respectively. All APPSwDI gene-modified mice were homozygous male mice.

In addition, in “cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice (n = 3)”, the blood concentration of this mouse was measured after administration of cilostazol and taxifolin-containing diet as described above. Or metabolites of cilostazol) and taxifolin (or metabolites of taxifolin) have been detected. Therefore, it has been confirmed that cilostazol and taxifolin are present in the mouse body.

In the Y maze test, the alternation response rate was measured from the results of the 8-minute test in accordance with a general implementation method. It is considered that the greater the change reaction, the better the working memory.

FIG. 7 is a graph showing the average turnover rate of each group of mice. Error bars represent standard error.

The alternation response rate was lower in APPSwDI gene-modified mice than in C57BL / 6J mice, suggesting impaired working memory. There was no significant improvement in the taxifolin-administered mice, but there was a tendency for the alternation response to increase in cilostazol and taxifolin-containing diet-administered APPSwDI gene-modified mice. Thus, it was shown that the combination of cilostazol and taxifolin can improve the impairment of working memory in APPSwDI genetically modified mice even when the dementia has progressed considerably.

Can be used for prevention and / or treatment of dementia.

Claims (6)

1. A carbostyril derivative comprising the following formula (1) (wherein R is a cycloalkyl group, A is a lower alkyl group, and a single bond or a double bond is present between the 3- and 4-positions of the carbostyryl nucleus. Or a salt thereof,
   

   

   
   Dihydroquercetin or a salt thereof;
   A drug for the prevention and / or treatment of dementia characterized by comprising:
2. A drug for the prevention and / or treatment of dementia,
   A carbostyril derivative having the following formula (1) as an active ingredient (wherein R is a cycloalkyl group, A is a lower alkyl group, and a single bond or a double bond is present between the 3- and 4-positions of the carbostyryl nucleus). A double bond) or a salt thereof,
   

   

   
   An active ingredient dihydroquercetin or a salt thereof;
   A drug for the prevention and / or treatment of dementia, administered in combination.
3. 3. The carbostyril derivative according to claim 1, wherein the carbostyryl derivative is 6- [4- (1-cyclohexyl-1H-tetrazol-5-yl) butoxy] -3,4-dihydrocarbostyril. A drug for the prevention and / or treatment of dementia.
4. The agent for preventing and / or treating dementia according to any one of claims 1 to 3, wherein the dihydroquercetin is taxifolin.
5. The dementia is Alzheimer's dementia, Lewy body dementia, frontotemporal dementia, cerebrovascular dementia, Parkinson's disease, Down's syndrome, or Huntington's disease. A drug for the prevention and / or treatment of dementia according to any one of the above.
6. The drug according to any one of claims 1 to 5, which is used for the prevention and / or treatment of peripheral symptoms of Alzheimer-type dementia.

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